JPS59155800A - Storable fluorescent sheet - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a stimulable phosphor sheet. More specifically, the present invention relates to a stimulable phosphor sheet that is particularly useful as a stimulable phosphor sheet for use in autoradiography using a measurement kit containing the stimulable phosphor sheet.

After administering a radioactively labeled substance to an organism, the organism or a part of its tissue is used as a sample, and this sample is overlapped with a radiographic film such as a high-sensitivity X-ray film for a certain period of time. Autoradiography (also called radioautography), which consists of exposing the film to light and obtaining positional information of the radiolabeled substance in the sample from the exposed area, has been known for a long time. ing. This autoradiography is used, for example, to study in detail the metabolism, absorption, and excretion routes and conditions of administered substances in living organisms. Such autoradiography is described, for example, in the following literature.

Biochemistry Experiment Course 6 Tracer Experiment Method (''-) 271~
289 pages, i'8. Autoradiography, 1 Toru Sueyoshi, Akiyo Shigematsu (1977, ■Tokyo Kagaku Doujin Series) Also, in recent years, autoradiography has been used to analyze tissue of an organism and/or substances derived from an organism that have been irradiated with a radioactive label. It can also be effectively used to obtain positional information of a radiolabeled substance in a support medium containing it.

For example, a radioactive label is added to a biologically derived polymeric substance such as a protein or a nucleic acid, and the radiolabeled polymeric substance, its derivative, or its decomposition product is subjected to separation and development operations such as gel electrophoresis. By developing the gel-like support medium for several minutes and overlapping the gel-like support medium and a high-sensitivity X-ray film for a certain period of time, the film is exposed to light. Methods for separating, identifying, or evaluating the molecular weight and characteristics of polymeric substances based on positional information have also been developed and are in actual use.

Such autoradiography is described, for example, in the following literature:

rELEcTROP)IOREsIs OF PRO
TETNS IN POLYACRYLAMTDE A
ND 5TARC: HGELSJ, A, H, Go
rdon (North-Holland Public
shing Company, Amsterdam
19f19): Japanese translation "Gel electrophoresis method," published by Tokyo Kagaku Dojin 1974.1 Especially in recent years, autoradiography has been
It has also been effectively used to determine the base sequence of nucleic acids such as, and is therefore an extremely useful means for determining the structure of macromolecular substances derived from living organisms.

By using this autoradiography, DN
Methods for determining the base sequence of A include 1, the Maxam-Gill/Hybert method, and the Sanger-Coulso method.
n) The law is known. In these methods, DNA has a double helical structure consisting of two chain molecules, and each of the two chain molecules contains four types of bases, namely adenine (A), guanine (G), It is composed of structural units containing the bases cytosine (C) and thymine (T), and the two chain molecules are cross-linked by hydrogen bonds between these four types of bases. A method of determining the base sequence by cleverly utilizing the characteristic structure of DNA, in which hydrogen bonds between each constituent nucleotide are realized only in two types of combinations, G-C and A-T. It is.

For example, the Maxam-in-Gilbert method is carried out by the operations described below.

DNA or DNA whose base sequence is to be determined
By bonding a group containing a radioactive isotope of phosphorus (P) to one end of a chain molecule of the decomposition product, the target substance is made into a radioactive labeling substance, and then using chemical means, Base-specific cleavage of bonds between each base in a chain molecule.

Next, the DNA obtained by this operation or a mixture of a large number of base-specific cleavage products of the DNA degradation product is separated and developed on a support medium by gel electrophoresis, and a large number of base-specific cleavage products are separated into bands. Minutes to form! You can get the expanded minute # expansion column (which cannot be seen visually). When this support medium and a high-sensitivity X-ray film are overlapped for a long time at low temperature, the part of the X-ray film facing the position where the base-specific cleavage product containing the radioactive isotope in the molecule is present. It is exposed to light and forms a latent image.

By developing the X-ray film with the latent image formed in this way, an autoradiograph consisting of a number of bands appears as a visible image on the X-ray film. and,
From this visualized autoradiograph and the specific cutting means, it is possible to sequentially determine the bases in a certain positional relationship from the end of the chain molecule to which the radioactive isotope is bound, and in this way, the target The sequence of all bases can be determined.

The Maxam-Gilbert method summarized in -1- is described in detail in the following document.

METHODS IN ENZYMOLOGY, V
OL, 85. PART I (ACADEMIC PR
ESS, NEW YORK LONDON
TR0NTO9YDNEY SAN FRAN
CISIGo, 1980) The Sanger-Cournon method is also a method that focuses on the characteristic structure of DNA and uses DNA synthase, gel electrophoresis, and autoradiography to determine the base sequence of DNA. - A detailed description of the characteristics and operation of the Cournon method and the aforementioned Maxam-Gilbert method can be found in the following documents:

"Reading genetic information in its original language: An unexpected DNA sequence analysis method," Kinbe Miura, Gendai Kagaku, September 1977 issue 4
Pages 6 to 54 (published by Tokyo Kagaku Dojin) As mentioned above, autoradiography is a process in which a sample consisting of a mixture of substances derived from an organism that has been labeled with a radioactive substance is transferred to a support medium (
For example, one-dimensional or two-dimensional position information of each substance separated and developed on the support medium after separation and development using a support medium for electrophoretic separation or support medium for thin layer chromatography. are effectively used to separate and identify substances by detecting and measuring them using their radioactivity. Therefore, by using this type of autoradiography, it is possible to efficiently determine, for example, the structure of biopolymers, and thus this type of autoradiography has been widely used in recent years.

However, there are several problems when actually utilizing such useful autoradiography.

First, in order to visualize the position of the radiolabeled substance separated and developed on the support medium, the support medium and high-sensitivity X-ray
The operation of exposing the film to light by overlapping it with a radiation film such as a line film for a certain period of time is complicated and requires a long time. That is, in conventional autoradiology, the above-mentioned exposure operation is performed at low temperatures (e.g., around 0°C, and -70 to -90°C in the case of exposing gel-like support media for nucleic acid base sequencing, etc.). , and is conducted over a long period of time (e.g., several days). This is because the usual radiolabeled substances that are filled in autoradiography generally do not have high radioactivity, so exposure must be long in order to obtain sufficient light sensitivity.
If a support medium and a radiation film are kept superimposed on each other for a long time at a relatively high temperature such as room temperature, the silver salt, which is a photosensitive component of the radiation film, will cause chemical fog due to various substances in the support medium. For this reason, it is difficult to obtain a highly accurate photosensitive image on the film, and therefore, the exposure operation must be carried out at a low temperature in order to reduce such chemical fog. In order to alleviate such harsh exposure conditions, it is possible to further increase the sensitivity of the radiation film, but conventional radiation films for autoradiography are already made with extremely high sensitivity. Considering the sharpness of the images obtained, it is difficult to dramatically increase the sensitivity of radiation films.

In addition, the silver salt of the photosensitive component of radiographic film has the disadvantage that it is easily affected not only by chemical stimuli but also by physical stimuli, which also makes autoradiography difficult to operate and reduces its accuracy. Become. That is, in autoradiography, it is generally necessary to perform the exposure operation with the radiation film in contact with a support medium, so operations such as moving and installing the radiation film are often performed with the radiation film in a bare state.

Therefore, during such work, there is an increased chance that the radiation film will come into contact with the operator's hands or equipment, etc.
Radiographic films tend to cause physical fog due to physical pressure caused by contact, etc., and this also causes a decrease in the accuracy of autoradiography. In order to avoid such physical fogging of the radiographic film, a high degree of skill and care is required in handling the film, which results in further complication of the autoradiography operation.

Furthermore, since conventional autoradiography involves long exposure operations as described above, the natural radioactivity contained in the sample in addition to the radiolabeled substance is also affected by the exposure of the radiographic film. There is a problem in that the accuracy of the location information of the radiolabeled substance is reduced. In order to eliminate such interference due to natural radioactivity, attempts have been made, for example, to conduct parallel experiments using control samples and to optimize the exposure time. The disadvantage is that the entire operation becomes complicated due to the necessity of preliminary experiments to determine the exposure time.

As a result of intensive research aimed at solving the above-mentioned problems associated with conventional autoradiography, the inventors of the present invention have discovered that stimulable fluorescent film can be used instead of radiation film as a photosensitive material for use in autoradiography. We have discovered that the above-mentioned problems can be solved or the drawbacks reduced by using a stimulable phosphor sheet having a phosphor layer made of a phosphor dispersed in a binder, and we have already filed a patent application for this invention ( Patent application 1934-18 (1982).

According to the autoradiography disclosed in the invention of the above-mentioned application, for example, the photosensitive material used to extract the positional information of the radiolabeled substance on the support medium for NF development where the sample is opened 11111 1. By using stimulable phosphor C-I, which has a phosphor layer made of a stimulable phosphor dispersed in a binder, it is possible to not only significantly shorten the exposure time, but also to reduce the environmental temperature. Alternatively, even under relatively high temperature conditions such as the temperature in the vicinity, exposure can be performed without reducing the accuracy of the obtained positional information of the radiolabeled substance. This point significantly simplifies the exposure operation in autoradiography, which has conventionally been carried out under cooling. Furthermore, by realizing a significant reduction in exposure time, the entire autoradiography operation can be carried out in a short time and efficiently, which is also very advantageous in practical use. Furthermore, by using the above-mentioned stimulable phosphor C-I as a photosensitive material for O-I radiography, chemical fog and physical fog, which have been a major problem when using conventional radiation films, are virtually eliminated. This also has a very advantageous effect on improving the accuracy and workability of radiography.

Furthermore, when a stimulable phosphor sheet is used as a photosensitive material, there is no need for special imaging in order to obtain positional information of the radiolabeled substance transferred to the stimulable phosphor sheet; By scanning the body sheet with electromagnetic waves such as a laser, the above position information can be read out, and the position information can be converted into an arbitrary form such as an image, symbol, numerical value, or a combination thereof. In general, by further processing the above position information using electrical means, it is also possible to obtain the necessary information in various desired forms.

Autoradiography using a stimulable phosphor sheet has various advantages as described above, and its usefulness is extremely high.

In the above autoradiography, for example,
A sample consisting of a mixture of substances from an organism in which a radioactive label is After separation and development, a stimulable phosphor sheet I can be superimposed on this support medium and an exposure operation can be carried out.However, the advantages of autoradiography using this stimulable phosphor sheet cannot be effectively utilized. In order to do this, a support medium is placed on the surface of the stimulable phosphor sheet in advance, and the sample is developed on the support medium in this state for several minutes, and if necessary, the support medium is separated and removed. , it is preferable to read the above-mentioned position information.

The work of installing the above-mentioned support medium on the surface of the stimulable phosphor sheet in advance involves, for example, applying an aqueous solution containing a polymer compound or its raw material components that are commonly used as constituent materials of the support medium to the stimulable phosphor sheet. C! It is carried out by coating and laminating on the second layer and then drying or heating. However, a protective film made of a hydrophobic plastic material is attached to the surface of a typical stimulable phosphor sheet, and the surface of this hydrophobic protective film is a component of the supporting medium. 1 or the aqueous solution of its raw material components, the problem is that it is difficult for the supporting medium formed on the optical sheet to adhere to the optical sheet. If the adhesion between the stimulable phosphor sheet and the support medium is poor, troubles such as detachment of the support medium, and especially voids between the stimulable phosphor sheet and the support medium, are likely to occur. The problem that the sample development operation performed on the support medium becomes non-uniform and, as a result, the accuracy of autoradiography is greatly reduced, tends to occur.

Therefore, it is not advantageous to use an ordinary stimulable phosphor sheet in order to perform autoradiography with the support medium previously provided on the stimulable phosphor sheet.

Therefore, in O-I radiography using a phosphor sheet as a photosensitive material, the present invention provides a support medium for developing a sample, or another support medium for containing a sample, using a stimulable phosphor sheet. The main object of the present invention is to provide a stimulable phosphor sheet which is particularly preferably used for carrying out an embodiment in which autoradiography is performed by pre-setting the sheet.

The present invention also provides a method for achieving high adhesion between the support medium and the stimulable phosphor sheet when performing an exposure operation by superimposing the support medium on which the sample has been separated and developed on the stimulable phosphor sheet. Another object of the present invention is to provide a stimulable phosphor sheet that can be used as a stimulable phosphor sheet.

The present invention comprises a support, a phosphor layer provided on the support and comprising a binder containing and supporting a stimulable phosphor in a dispersed state, and a protective film provided on the phosphor layer. The stimulable phosphor sheet is characterized in that the surface of the protective film is imparted with hydrophilic properties.

Next, the present invention will be explained in detail.

Typical embodiments of the stimulable phosphor sheet of the present invention include:
The following stimulable phosphor sheets can be mentioned.

(1) a support, a phosphor layer provided on the support, a phosphor layer comprising a binder supporting the exhaustible phosphor in a dispersed state, and a protective layer provided on the phosphor layer; A stimulable phosphor sheet consisting essentially of a film,
An 87-laminated phosphor sheet whose protective film has been treated with wood to make it hydrophilic.

(2) A TLF light layer consisting of a support, a binder containing and supporting the stimulable phosphor provided on the support in a dispersed state, and the ? ``A stimulable phosphor sheet substantially consisting of a protective film provided on a phosphor layer,
A stimulable phosphor sheet that has a hydrophilic layer on top of a protective film, making it hydrophilic.

Substantially consisting of a support, a phosphor layer provided on the support and made of a binder containing and supporting a stimulable phosphor in a dispersed state, and a protective film provided on the phosphor layer. The stimulable phosphor sheet disclosed in the US Pat. No. 3.8 is also called a radiation image conversion panel.
59.527 and Japanese Patent Laid-Open No. 55-12145, etc., and the general structure is already known.

6 In other words, the stimulable phosphor sheet absorbs the radiation energy transmitted through the subject or the radiation energy emitted from the subject into the stimulable phosphor of the panel, and then exposes the stimulable phosphor to visible light and By exciting the stimulable phosphor in a time-series manner using electromagnetic waves (excitation light) such as infrared rays, the radiation energy stored in the stimulable phosphor is released as fluorescence, and this ILF light is read photoelectrically to generate an electrical signal. This electrical signal is reproduced as a visible image on a recording material such as a photosensitive film, a display device such as a CRT, or is expressed as numerical or symbolic position information.

The stimulable phosphor C I, which is preferably used in the present invention, will be briefly explained below.

The phosphor layer that provides the basic functions of the stimulable phosphor sheet is composed of a stimulable phosphor and a binder that contains and supports the stimulable phosphor in a dispersed state. After absorbing radiation, it has the property of emitting light (stimulated luminescence) when irradiated with electromagnetic waves (excitation light) such as visible light and infrared rays. Therefore, for example, radiation emitted from an object such as a sample containing a radiolabeled substance is absorbed by the phosphor layer of the stimulable phosphor sheet in proportion to the amount of radiation, and the radiation is absorbed by the phosphor layer of the stimulable phosphor sheet. A radiation image of the subject is formed as an image of accumulated radiation energy. This accumulated image is generated by electromagnetic waves such as visible light and infrared light (
By excitation with excitation light), stimulated luminescence (fluorescence) can be emitted, and by photoelectrically reading this stimulated luminescence and converting it into an electrical signal, the accumulated image of radiation energy can be converted into a visible image or It becomes possible to convert into numerical values, symbols, etc. indicating the location information of radioactive (labeled) substances.

The support for the stimulable phosphor sheet can be arbitrarily selected from various materials used as supports for intensifying screens in conventional radiography. Examples of such materials include films of plastic materials such as cellulose acetate, polyester, polyethylene terephthalate, polyamide, polyimide, triacetate, polycarbonate, metal sheets such as aluminum foil, aluminum alloy foil, ordinary paper, Examples include baryta paper, resin-coated paper, pigment paper containing pigments such as titanium dioxide, and paper sized with polyvinyl alcohol. However, when considering the characteristics and handling of the stimulable phosphor sheet as an information recording material, a particularly preferred material for the support in the present invention is a plastic film. A light-absorbing substance such as carbon black may be incorporated into this plastic film.
Alternatively, a light-reflecting substance such as titanium dioxide may be incorporated. The former is a support suitable for a high sharpness type stimulable phosphor sheet, and the latter is a support suitable for a high sensitivity type stimulable phosphor sheet.

In known stimulable phosphor sheets, in order to strengthen the bond between the support and the phosphor layer, or to improve the sensitivity or image quality of the stimulable phosphor sheet, the surface of the support on the side where the phosphor layer is provided is A polymeric substance such as gelatin is coated on the surface to form a zero-adhesive layer, or a light-reflecting layer made of a light-reflecting substance such as titanium dioxide or a light-absorbing layer made of a light-absorbing substance such as carbon black is provided. This is also being done. The support used in the present invention can also be provided with these various layers, and their configurations can be arbitrarily selected depending on the desired purpose, use, etc. of the stimulable phosphor sheet.

Furthermore, as disclosed in Japanese Patent Application No. 57-82431 filed by the present applicant, in order to improve the sharpness of the resulting image, the surface of the support on the phosphor layer side (the phosphor layer side of the support) When an adhesion-imparting layer, a light-reflecting layer, a light-absorbing layer, a metal foil, or the like is provided on the surface of the layer, the surface (meaning the surface) may be provided with irregularities.

A phosphor layer is formed on the support as described above. The phosphor layer is basically a layer made of a binder containing and supporting particulate stimulable phosphor in a dispersed state.

As mentioned above, a stimulable phosphor is a phosphor that exhibits stimulated luminescence when irradiated with radiation and then irradiated with excitation light.
From a practical standpoint, a phosphor that exhibits stimulated luminescence in a wavelength range of 300 to 500 nm by excitation light in a wavelength range of 400 to 800 nm is desirable. The stimulable phosphor used in the stimulable phosphor sheet used in the present invention is preferably a divalent europium-activated alkaline earth metal fluorohalide phosphor, but is not limited thereto. It's not something you can do.

Examples of the stimulable phosphor used in the stimulable phosphor sheet of the present invention include SrS:Ce, Sm, and SrS:Eu, which are described in US Pat. No. 3,859,527.

Sm, Th02: Er, and La2o2s: Eu
, a phosphor expressed by a composition formula such as Sm, JP-A-55
ZnS: Cu, P described in -12142 publication
b, BaO*xAl2O3:Eu [however, 0.8≦X
≦10], and MπO@xsi02 :A [where M■ is Mg, Ca, Sr, Zn, Cd, or Ba
and A is Ce, Tb, Eu, Tm, Pb, TI, B
i, or Mn, and X is a phosphor represented by a composition formula such as 0.5≦X≦2.5, JP-A-55-1
It is described in Publication No. 2143 (B at-X
-)', Mgx, Cay) FX
:aEu” [However, X is at least one of 0 sentence and Br, and X and y are 0 and x+y≦0
．． 6, and x y#o, and a is 10-6≦a≦5
A phosphor represented by the composition formula of
X: xA [where Ln is La, Y, Gd, and Lu
X is at least one of C1 and Br, A is at least one of Ce and Tb, and X is O<x<0.1. The expressed phosphor is described in Japanese Unexamined Patent Publication No. 12145/1983 (B
al-) (, M” One, A is Eu, Tb, Ce, Tm, Dy
, Pr, Ho, Nd, Yb, and Er, and X is 0≦X≦o, e, and y are 0≦y.
≦0.2] A phosphor represented by the composition formula M" described in JP-A No. 55-160078
FX-xA: yLn [However, Mll is Ea, C
at least one of a, Sr, Mg, Zn, and Cd, A is BeO, MgO1CaO1SrO, BaO
lZnOlA sentence 203. Y2O3, La2O3, In2
O3,5i02, TM01, Z r02, GeO2,5
n02, Nb2O5, Ta205, and ThO2, Ln is Eu, Tb, Ce, Tm
, Dy, Pr, Ho, Nd, Yb, Er, Sm
, and at least one of Gd, X is t, Q, B
r, and at least one of ■, and X and y are respectively 5
<4<0.2, a phosphor represented by the composition formula of
at-x, M”X)F1a aBaX2:yEu,
zAE However, Mm is at least one of beryllium, magnesium, calcium, strontium, zinc, and cadmium, X is at least one of chlorine, bromine, and iodine, and A is at least one of zirconium and scandium. Yes, a, x, y, and 2 are each 0.5≦a≦1.25.0≦X≦1.
10-@≦y<2X10-' and O<z≦10-2
A phosphor represented by the composition formula, JP-A-57-2
Described in Publication No. 3673 (B al-)
(, M”X) F 2 ” aB a
X2: yEu, zB [However, MI [represents helium, magnesium, calcium, strontium, zinc,
and at least one of cadmium, X is chlorine,
at least one of bromine and iodine, a,
x, y, and 2 are each 0.5≦a≦1.25, o
≦X≦1.10'<y<2X10-1, and O<z≦
A phosphor represented by the composition formula 2X10-"t'" is described in Japanese Patent Application Laid-Open No. 57-23675 (
B a 1-X, M''X
) F 2 ・ a B a X 2
:yEu, zA [However, MU is at least one of beryllium, magnesium, calcium, strontium, zinc, and cadmium, X is at least one of chlorine, bromine, and iodine, and A is at least one of arsenic and silicon. a, x, y, and Z are each 0.5≦a≦1,25, O≦X≦1.1
0′−”≦y≦2×10−I, and 0<z≦5 X
A phosphor represented by the composition formula M"OX:xCe [where M is Pr, Nd, Pm, Sm, Eu, Tb, Dy, Ho, E
is at least one trivalent metal selected from the group consisting of r, Tm, Yb, and Bi, X is one or both of C1 and Br, and X is O<
A phosphor represented by the composition formula: B a + -x M x /2 L x / described in Japanese Patent Application No. 1989-89875 filed by the present applicant.
2 F X :yEu'' [However, 1M is Li, Na,
represents at least one alkali metal selected from the group consisting of Rh and Cs; L represents Sc, Y, La
, Ce, Pr, Nd, Pm, Sm, Gd, Tb, Dy,
Ho, Er, Tm, Yb, Lu, AfL, Ga, In,
and at least one kind of trivalent metal selected from the group consisting of Ti; X represents at least one kind of halogen selected from the group consisting of O, Br, and ■; 0.5, y is o<y≦0,1
BaFXkenxA:yEu2+ [where X
is at least one kind of halogen selected from the group consisting of C1, Br, and A; A is a fired product of a tetrafluoroboric acid compound; and X is 101≦X≦
o, t, y are O7 y≦0.1] A phosphor represented by the composition formula BaFX* xA: yEu” [ However, X is at least one kind of halogen selected from the group consisting of C1, Br, and E; A is a monovalent or divalent metal salt of hexafluorosilicon, hexafluorotitanic acid, and hexafluorozirconium is a fired product of at least one compound selected from the hexafluoro compound group consisting of; and X is 10-'≦X≦0
゜1, y is o<y≦0.1] A phosphor represented by the composition formula BaFX@xNaX':aEu''[ However, X and X゛ are each at least one of 0 sentence, Br, and ■, and X and a are each O<
x≦2, and Ova≦0.2] A phosphor represented by the composition formula: 8 M”F X':
yEu": zA [where MlX is at least one alkaline earth metal selected from the group consisting of Ba, Sr, and Ca; X and X' are C1, B, respectively
r, and at least one kind of halogen selected from the group consisting of I; A is V, Cr, Mn, Fe, Co
, and at least one transition metal selected from Ni; and X is O<x≦2, y is o<y≦0.2,
and 2 is 0<z≦10-2], M"FX・aM '
“X” 2 * cm Sword “X”
3 Fists xA: yEu 2+ [However, M
M is at least one alkaline earth metal selected from the group consisting of Ba, Sr, and Ca, and M x is L
is at least one alkali metal selected from the group consisting of i, Na, Rb, and Cs; M'1 is Be
and Mg, and M III is A! ;L, Ga, In,
and at least one trivalent metal selected from the group consisting of Tu; A is a metal resistant material;
, and X′° are at least one halogen selected from the group consisting of F, 0, Br, and ■; and , a is O≦a≦2. b is O≦b≦10-2, C is O≦C≦1O-2, and a+b
+c≧1O−1l; X is O<x≦0.5, y is o
Examples include a phosphor represented by the composition formula <y≦0.2].

However, the stimulable phosphor used in the present invention is not limited to the above-mentioned phosphors, but any phosphor that exhibits stimulated luminescence when irradiated with radiation and then excitation light. It may be something.

Examples of binders for the optical layer include proteins such as gelatin, polysaccharides such as dextran, or natural polymeric substances such as gum arabic; and polyvinyl butyral, polyvinyl acetate, nitrocellulose,
Ethyl cellulose, hnylidene chloride/vinyl chloride copolymer, polymethyl methacrylate, vinyl chloride/vinyl acetate copolymer, polyurethane, cellulose acetate!
- Binders typified by synthetic polymeric substances such as butyrate, polyvinyl alcohol, linear polyester, etc. can be mentioned. Particularly preferred among such binders are nitrocellulose, linear polyesters, and mixtures of nitrocellulose and linear polyesters.

The phosphor layer can be formed on the support, for example, by the following method.

First, the above stimulable phosphor particles and binder are mixed in a suitable solvent (
For example, a lower alcohol, a chlorine atom-containing hydrocarbon, a ketone, an ester, an ether) are added to the binder solution and thoroughly mixed to prepare a coating solution in which phosphor particles are uniformly dispersed in a binder solution.

The mixing ratio of the binder and the stimulable phosphor particles in the coating solution varies depending on the characteristics of the desired stimulable phosphor sheet, the type of phosphor particles, etc., but in general, the mixing ratio of the binder and the stimulable phosphor particles is The mixing ratio is selected from the range of 1:1 to 1.100 (by weight), and especially 1:8 to 1:40.
(weight ratio).

The coating IIj liquid contains a dispersant to improve the dispersibility of the phosphor particles in the coating liquid, and a bonding force between the binder and the phosphor particles in the phosphor layer after formation. Various additives such as plasticizers for improving the properties may be mixed. Examples of dispersants used for such purposes include phthalic acid, stearic acid, capronic acid, lipophilic surfactants, and the like. Examples of plasticizers include triphenyl phosphate, tricresyl phosphate,
Phosphate esters such as diphenyl phosphate; phthalate esters such as diethyl phthalate and dimethoxyethyl phthalate;
Glycolic acid esters such as ethyl phthalyl ethyl glycolate and butyl phthalyl glycolate; and polyesters of polyethylene glycol and aliphatic compounds such as polyesters of triethylene glycol and adipine, and polyesters of diethylene glycol and succinic acid. Examples include polyester with basic ugliness.

The coating solution containing the phosphor particles and binder prepared as described in Section 2 is then uniformly applied to the surface of the support to form a coating film of the coating solution. This coating operation can be carried out using conventional coating means such as a doctor blade, roll coater, knife coater, etc.

The formed coating film is then dried by gradually heating to complete the formation of the phosphor layer on the support. The thickness of the phosphor layer varies depending on the characteristics of the desired stimulable phosphor, the type of phosphor particles, the mixing ratio of the binder and the phosphor particles, etc., but is usually between 20 JLm and 1 mm. However, this layer thickness is preferably 50 to 500 JLm.

Note that the phosphor layer does not necessarily need to be formed by directly applying the coating liquid onto the support as described above; for example, the phosphor layer may be formed by separately applying the coating liquid to a sheet such as a glass plate, metal plate, or plastic sheet. After the phosphor layer is formed by coating and drying, it may be pressed onto the support, or the support and the phosphor layer may be joined by a method using an adhesive.

A protective film is provided on the fit light layer as described above. This protective film is made of, for example, transparent cellulose derivatives such as cellulose acetate and nitrocellulose; polymethyl methacrylate, polyvinyl butyral, polyvinyl formal, polycarbonate, polyvinyl acetate/vinyl acetate copolymer, polyethylene terephthalate, polyethylene, polyvinylidene chloride, polyamide, etc. It is made of a transparent synthetic polymer material. The thickness of the protective film is usually 1 to 100 pm, preferably 3 to 50 km.
It is said that

In the present invention, hydrophilicity is imparted to the surface of the protective film of the stimulable phosphor sheet formed as described below.

Typical methods for making the surface of the protective film of a stimulable phosphor sheet hydrophilic include a method of treating the surface of the protective film to make it hydrophilic, and a method of providing a hydrophilicity-imparting layer on the protective surface.

Examples of the former method of making the surface of the protective film hydrophilic include a method using the following surface activation treatment. Namely, chemical treatment with chemicals such as acid, alkali, and etching solution; physical treatment such as roughening treatment; electrical treatment with corona discharge, high frequency discharge, glow discharge, active plasma, etc.; and light treatment with ultraviolet rays, laser, etc. Examples include treatment; flame treatment; ozone oxidation treatment.

In the latter method of providing a hydrophilicity-imparting layer on the surface of the protective film, examples of materials used for the hydrophilicity-imparting layer include natural polymeric substances and their derivatives such as gelatin, starch, agarose, and cellulose; polyvinyl alcohol, polyvinylpyrrolidone; Synthetic homopolymers such as polyacrylamide, polyhydroxyethyl methacrylate, etc.; Wood-friendly synthetic polymer substances such as synthetic copolymers obtained by copolymerization with ethylenically unsaturated monomers such as esters, acrylic acid esters, vinyl chloride, and vinylidene chloride, and dienes such as butadiene, isoprene, and inbutylene. can be mentioned.

The above-mentioned hydrophilicity-imparting layer may be placed on the protective film by a known layer forming method, such as applying it to the sheet surface as a solution dissolved in water or other solvents, or as a latex-like dispersion. I can do it. Note that the wood-philicity imparting layer may be a single layer or may be laminated as a plurality of layers.

When carrying out a method of imparting hydrophilicity to a protective film of a stimulable phosphor sheet, for example, US Pat.
698, No. 241, No. 2,764,520, No. 2.
No. 864.755, No. 2, 864.

No. 756, No. 2.972.534, No. 3,056 7.792, No. 3.071.466, No. 3゜072
．． No. 483, No. 3,143,421, No. 3.145,
No. 105, No. 3.145.242, No. 3.360.4
No. 48, No. 3,376.208, No. 3.462.33
No. 5, No. 3,475°193, British Patent No. 788.3
It is possible to refer to methods for making the surface of plastics hydrophilic, which are described in Japanese Patent No. 65, No. 804.005, No. 891,489, and the like.

When the stimulable phosphor sheet of the present invention is used as a component of an autoradiography measurement kit, the other component, the support medium, that is, the biologically-derived substance labeled with a radioactive label, is separated and developed. A measurement kit is provided as a supporting medium for the measurement on the protective surface of the stimulable phosphor sheet.

The above-mentioned support medium can be arbitrarily selected from various support media for separation and development that are used or proposed for use in conventional autoradiography techniques. Examples of such support media for separation development include support media for electrophoretic separation in the form of gel support media, polymer moldings such as acetate membranes, or various support media such as filter paper, and silica gel. Mention may be made of support media for thin layer chromatography.

Note that the support media that can be used are not limited to those exemplified above, but any support medium that can be used for separation and development of samples in autoradiography technology and that can be attached to a stimulable phosphor sheet. Any support medium can be used.

It is preferable that the support medium for separation and development is applied onto the stimulable phosphor sheet at the same time as the support medium is formed. That is, it can be carried out according to a conventional method in the same manner as when forming a support medium on a normal substrate (supporting aid) such as a glass plate or a plastic film. for example,
After dissolving (or dispersing) the support medium raw material in an appropriate solvent to form a solution (or suspension), this solution is applied to the surface of the protective film of the stimulable phosphor sheet, and a plastic frame is placed on the sheet. The support medium can be formed, such as by pouring into it or simply coating it. However, the support medium can also be formed in advance on a common substrate such as a glass plate or a plastic film according to a conventionally known technique, and then provided on the surface of the protective film of the stimulable phosphor sheet.

The solvent to be used can be arbitrarily selected depending on the purpose from various known solvents, pH-adjusted buffers, etc. in combination with the support medium raw materials. Such solvents are already well known and will not be specifically mentioned here.

Next, an autoradiography operation using a measurement kit using the stimulable phosphor sheet of the present invention will be described.

Examples of samples to be separated and developed, that is, radioactively labeled biological substances, include proteins, nucleic acids,
Examples include polymeric substances such as derivatives thereof and decomposition products 9 thereof. Methods for imbuing these substances with radioactive labels are already well known. Note that the biologically derived substances to be measured by the autoradiography measurement kit are not limited to the above-mentioned polymeric substances.

In addition, separation and development methods such as electrophoresis are carried out using the various support media for separation and development as described above, and a method in which a sample is developed on the support medium is also well known. I don't really touch it.

Next, by irradiating the autoradiography measurement kit with the sample separated and developed on the support medium with appropriate light, heat, etc., if necessary, the stimulable phosphor sheet accumulates in the sample separation and development process. Radiation energy is emitted as fluorescence. In other words, the measurement key is determined by the natural radioactivity contained in the sample, and by the radiation emitted from the moving radiolabeled substance during the process in which the radiolabeled sample is separated and developed on a support medium. When the stimulable phosphor sheet of the kit is exposed to light, a radiation energy accumulation image other than the object to be measured is formed on the stimulable phosphor sheet, which causes noise and noise to the radiation energy accumulation image containing the target autoradiograph. Become. Therefore, if the influence of the noise is not negligible, it is desirable to eliminate the noise before forming a radiation energy accumulation image having the desired autoradiograph on the stimulable phosphor sheet.

The above-mentioned noise erasing operation can be performed on the support medium on which the sample has been separated and developed, or after it has been subjected to any desired treatment such as drying or fixation of the separated and developed product.

Next, place the autoradiography measurement kit in which the sample has been separated and developed on the attached support medium for separation and development.
Preferably, the stimulable phosphor sheet is exposed to light by leaving it in a dark place or in a dark box for a certain period of time, and at least a portion of the radiation emitted from the separated and developed radioactive label substance is absorbed into the stimulable fluorescence in the measurement kit. Let it be absorbed into the body sheet.

Through this operation, the autoradiograph of the radiolabeled substance separated and developed on the support medium is transferred to the stimulable phosphor sheet as an image of accumulated radiation energy.

In the above exposure operation, the exposure time varies depending on the strength of the radioactivity of the radiolabeled substance contained in the sample, the concentration and density of the substance, or the sensitivity of the stimulable phosphor sheet. However, when a stimulable phosphor sheet is used as the photosensitive material, the exposure time is significantly shortened compared to the exposure time required when a conventional radiation film is used.

In addition, in the operation of reading the positional information of the radiolabeled substance on the support medium that has been transferred and accumulated from the support medium to the stimulable phosphor sheet by exposure, the intensity and distribution of the energy stored in the phosphor sheet can be measured. , it is possible to change the state of the obtained position information by performing various electrical processes depending on the desired information, etc., so there is no particular need to strictly control the exposure time during the exposure operation.

There is no particular restriction on the temperature at which the exposure operation is performed, but the temperature at which the stimulable phosphor sheet of the present invention is used is not limited. - Radiography can be carried out at ambient temperatures, especially from 10 to 356C. However, lower temperatures such as those utilized in conventional autoradiography (e.g.
The exposure operation may be performed at a temperature of about 5° C. or lower.

Next, the stimulable phosphor sheet on which the autoradiograph of the sample has been transferred and accumulated is subjected to a reading process. The readout process can be performed with the support medium attached to the M-laminated phosphor sheet, or after the stimulable phosphor sheet is separated from the measurement kit, but only the latter stimulable phosphor sheet can be read out. It is desirable to apply it to the process.

The support medium for separation and development can be easily removed from the measurement kit by, for example, peeling or scraping the support medium, or washing it away with a solvent such as water, depending on the purpose. .

In the present invention, the method for reading out the positional information of the radiolabeled substance 3 possessed by the autoradiograph transferred and accumulated on the stimulable phosphor sheet is described using the reading device (or reading device) shown in FIG. 1 of the accompanying drawings. This will be briefly described below with reference to an example.

Figure 1 shows a readout system for reading out one-dimensional or two-dimensional positional information of a radiolabeled substance accumulated and recorded on a stimulable phosphor sheet (hereinafter sometimes abbreviated as phosphor sheet). 1 shows a schematic diagram of an example of an apparatus.

In the reading device, the following reading operation is performed.

By passing the laser beam 3 generated from the laser light source 2 through the filter 4, a portion of the wavelength region corresponding to the wavelength region of stimulated luminescence generated from the phosphor sheet 1 in response to excitation by the laser beam 2 is cut off. Ru. The beam diameter of the laser beam 2 that has passed through the filter 4 is then precisely adjusted by a beam expander 5. Next, the laser beam is deflected by a light deflector 6 such as a galvanometer mirror, reflected by a plane reflecting mirror 7, and then one-dimensionally deflected and incident on the phosphor sheet 1 4 . Note that an fθ lens 8 or the like is arranged between the optical deflector 6 and the plane reflecting mirror 7, so that when the deflected laser beam scans the phosphor sheet 1, a uniform beam speed is always maintained. ing.

The laser light source 2 used here is selected so that the wavelength range of its laser light 4 does not overlap with the main wavelength range of stimulated luminescence emitted from the phosphor sheet 1.

The phosphor sheet 1 is transported in the direction of the arrow 9 under irradiation with the above-mentioned polarized laser light. Therefore, the entire surface of the phosphor sheet 1 is irradiated with the polarized laser light.

When the phosphor sheet 1 is irradiated with the laser light as described above, it exhibits stimulated luminescence with an amount of light proportional to the radiation energy stored and recorded, and this light enters the light guide sheet 10. The light guide sheet 10 has a linear incident surface and is placed close to the scanning line on the phosphor sheet 1 so as to face it, and its exit surface forms an annular ring. It is connected to the light receiving surface of the photodetector 11. The light guide sheet 10 is made by processing a transparent thermoplastic resin sheet such as acrylic synthetic resin, and allows light incident from the incident surface to be transmitted to the exit surface while being totally reflected inside. It is configured to The stimulated luminescence from the phosphor sheet 1 is guided through the light guide sheet 10 and reaches the exit surface, is emitted from the exit surface, and is received by the photodetector 11.

The preferred shape, material, etc. of the light-guiding sheet are disclosed in Japanese Patent Application Laid-open No. 5
Disclosures are made in JP-A No. 5-87970, JP-A No. 56-11397, and the like.

A filter is attached to the light-receiving surface of the photodetector 11, which transmits only light in the wavelength region of stimulated luminescence and cuts light in the wavelength region of excitation light (laser light), and detects only stimulated luminescence. It is made to be wet. The stimulated luminescence detected by the photodetector 11 is converted into an electrical signal, and the sensitivity is set in the amplifier 1 according to the amplification factor setting value a output from the control circuit 12.
After being amplified to an appropriate level of electrical signal in step 3, A
/D converter 14. The A/D converter 14 converts the digital signal into a digital signal with a scale factor suitable for the signal fluctuation width in accordance with the recording scale factor setting value also output from the control circuit J2, and inputs the digital signal to the signal processing circuit 15. The signal processing circuit 14 performs signal processing based on the reproduced image processing condition setting value C also output from the control circuit 12 so as to obtain an excellent visible image with appropriate density and contrast and suitable for observation and interpretation. Then, if necessary, the data is electronically transmitted to a recording device (not shown) via a storage means such as a magnetic tape.

Note that the amplification factor setting value a, recording scale factor b, and reproduction image processing condition setting value C output from the control circuit 12 are determined by, for example, performing a preliminary reading operation (pre-reading operation) before the above-mentioned reading operation. The factors a, b, and c can be set so that an image with the most uniform density and contrast and excellent observation and interpretation performance can be obtained according to the accumulated recorded information obtained by performing the above steps, or If the content of radioactive substances in a sample is known in advance, the number of stimulable phosphor 7 sheets for that sample can be determined empirically.

Recording devices include, for example, those that optically record by scanning a photosensitive material with a laser beam, etc., those that electronically display on a CRT, etc., and those that display X-ray images displayed on a CRT, etc. on a video printer, etc. Recording apparatuses based on various principles can be used, such as those that record information on heat-sensitive recording materials using heat rays, and those that record on heat-sensitive recording materials using heat rays.

However, the recording device is not limited to one that creates a visible image as described above, but also one that converts the one-dimensional or two-dimensional positional information of the radiolabeled substance in the sample into numbers or symbols, as described above. You can also record it by doing something like this.

Furthermore, as a method for reading the positional information of the radiolabeled substance contained in the autoradiograph transferred and stored on the stimulable phosphor sheet, it is of course possible to use any method other than those exemplified above.

In addition, in this specification, "position information" of radiolabeled substances in a sample refers to various types of information centered on the position of radiolabeled substances or their aggregates in a sample, such as Refers to various types of information obtained as one or any combination of information such as the location and shape of an aggregate of radioactive materials, the concentration and distribution of radioactive materials at that location, etc.

In addition, in the descriptions so far in this specification, as a particularly useful embodiment of the hydrophilic stimulable phosphor sheet of the present invention, a radioactive label is added to the surface of the protective film of the stimulable phosphor sheet. Although an example has been shown in which a support medium for separating and developing a sample is stabbed and autoradiography is performed using this, the embodiment of the stimulable phosphor sheet of the present invention is not limited to this. For example, when attaching an arbitrary hydrophilic layer such as a culture medium for microorganisms to a stimulable phosphor sheet, the stimulable phosphor sheet provided with wood-philicity according to the present invention can be advantageously used.

Note that the stimulable phosphor sheet of the present invention does not necessarily need to be impregnated with a support medium for sample separation and development, a culture medium, or the like in advance. That is, in order to obtain positional information of a radiolabeled substance in a support medium on which separation and development of a sample containing a radiolabeled substance has been completed, or in a medium in which a sample containing a radiolabeled substance has been cultured, the support medium, etc. Even in the case where the exposure operation is carried out by overlapping, the present invention is useful because it has high adhesion to the supporting medium, which simplifies the exposure operation and makes it possible to improve the accuracy of the positional information obtained. be.

Next, an embodiment of autoradiography using the measurement kit using the stimulable phosphor sheet of the present invention will be described, taking as an example the initial operation of the DNA base sequencing method using the aforementioned Maxam-Gilbert method. .

[Example 1] Stimulable europium-activated barium fluoride bromide phosphor (B
Methyl ethyl ketone was added to the mixture of particles of aFBr+Eu) and linear polyester resin, and the nitrification degree was 11.
．． A dispersion containing phosphor particles in a dispersed state was prepared by adding 5% nitrocellulose. Next, tricresyl phosphate, n-butanol, and methyl ethyl ketone were added to this dispersion, and then sufficiently stirred and mixed using a propeller mixer to ensure that the phosphor particles were uniformly dispersed and the viscosity was 25 to 35 PS (25 A coating solution was prepared.

Carbon black kneaded polyethylene terephthalate sheet placed horizontally on a glass plate (support, thickness: 25
This coating liquid was applied uniformly onto the surface of the sample (0ILm) using a doctor blade. After coating, the support on which the coating film was formed was placed in a dryer, and the temperature inside the dryer was gradually raised from 25° C. to 10,000° C. to dry the coating film.

In this way, a phosphor layer having a layer thickness of 300 #Lm was formed on the support.

Next, on top of this phosphor layer, a polyester adhesive was applied to one side of a transparent polyester terephthalate film (CW size: 12 pLm), and the film was bonded by placing the adhesive layer side facing down. A protective film was formed to obtain a stimulable phosphor sheet composed of a support, a phosphor layer, and a protective film.

Next, glow discharge treatment was performed on the surface of the protective film of the stimulable phosphor sheet. This glow discharge treatment is performed at 0.05 mmH.
A semicircular rod-shaped electrode (
Diameter of cross section: 2cm, length: 40cm) 4 trees are 10cm
The protective film of the stimulable phosphor sheet was applied at a position 15 cm apart from the electrode plate, which was installed on an insulating plate at intervals, under conditions of a discharge voltage of 3 kV, a processing interval of 3 seconds, and an electrode current of 0.4 A. This was done by operating the robot so that it ran opposite the electrode surface.

Then, a stimulable phosphor sheet was placed on a 4 mm thick glass plate with the glow discharge treated protective film facing upward, and a rectangular polymethyl methacrylate spacer (thickness:
1.5mm) to create a frame, sandwich it between glass plates of the same thickness, and place a mold (1.5mm x 20mm) between this glass plate 2 and the phosphor sheet.
0mm x 200mm) was removed. During this mold,
A tris-housing acrylamide buffer (acrylamide concentration = 8%, crosslinking agent ratio: 3%) prepared by a conventional method was injected and polymerized.

In this way, a measurement kit in which a slab gel of polyacrylamide was formed was obtained.

(2) Autoradiograph - Escherichia coli plasmid DNA (pBR322) by standard method
After cutting with the restriction enzyme Hind-H, the 5'-end was labeled with 32F to obtain double-stranded DNA (32P-labeled product).
1 hong was obtained.

Separately prepared 20 mM Tris[tris(hydroxymethyl)aminomethane]/hydrochloric acid buffer (p
H7, 4) Add the above double-stranded DNA I and about 1 unit of the restriction enzyme Hae-N to 20 g of the fragment, perform a specific decomposition reaction at 37°C for 1 hour, and remove the decomposition products of the above fragments. A decomposition mixture solution was obtained.

Using this decomposition mixture solution as a sample, the measurement temperature described above]
and using 50mM Tris-borate buffer (pH 8.3) containing 1mM EDTA as the electrode solution, a voltage of 5.
Electrophoresis operations were carried out on slab gel supports at 00V. The electrophoresis was stopped when the marker dye previously added to the sample reached the lower end of the gel, and the origin of the coordinate axes was marked with 32F-containing ink.

Next, the measurement kit described in "2" was exposed to light to erase noise on the stimulable phosphor sheet, and then left in a dark box at room temperature (approximately 25° C.) for 1 minute to carry out the exposure operation.

After taking out only the stimulable phosphor sheet from this measurement kit in a dark room, introduce the stimulable phosphor sheet into a reading device as shown in Figure 1, and place the mark with the 32F-containing ink on the coordinate axis. As the origin, positional information indicating the migration position of the degradation product of the 32P-labeled fragment was read. Then,
According to this positional information, a gel portion containing a 32F-labeled decomposition product from the separated slab gel support was cut out using a thin razor, and this was transferred to a test tube.

For confirmation, the remaining gel that had been partially excised as described in section 2 was superimposed on the stimulable phosphor sheet again, and then a readout device was used to check whether or not any decomposition product with a 32P label remained. Upon examination, it was found that the entire amount of the 32p-labeled decomposition product was removed. That is, it was confirmed that the positional information of the decomposition product having the 32p label obtained through the stimulable phosphor sheet described in "2" was highly accurate.

[Brief explanation of the drawing]

Figure 1 shows an example of a reading device (or reading device) for reading the positional information of a radiolabeled substance in a sample that has been transferred and accumulated on a stimulable phosphor sheet in autoradiography using a stimulable phosphor sheet. This shows that. 1: stimulable phosphor sheet, 2: laser light source, 3: laser light, 4: filter, 5: beam expander, 6: light deflector, 7: plane reflecting mirror, 8: fθ lens, 9
: Transfer direction, 10: Light-guiding 5 sheet, 11: Photodetector, 12: Control circuit, 13: Amplifier, 14: A/D converter, 15 = Signal processing circuit Patent applicant Fuji Photo Film Co., Ltd. Agent Patent Attorney Yasuo Yanagawa 6 Procedural Amendments Patent Office Commissioner Kazuo Wakasugi], Case Description 1981 Patent Application No. 30605 2° Title of Invention Stormative Phosphor Sheet 3
Relationship with the case of the person making the amendment: Patent applicant 4, Agent 6, Number of inventions increased by the amendment: None 7, “Detailed description of the invention” column 8 of the specification subject to the amendment, Contents of the amendment enM-tr+Jr +-4y /7'□The "Detailed Description of the Invention" column of the specification will be amended as follows. Kiichiyu Fukumachi Jingni - Correction Ikuichi (1) Page 17, line 17 U.S. Patent No. 3,859. → Deletion ~ Line 18 of the same page Specification No. 527 and above

Claims (1)

[Claims] l. The phosphor layer consists essentially of a support, a phosphor layer provided on the support that contains and supports a stimulable phosphor in a dispersed state, and a protective film provided on the phosphor layer. 1. A stimulable phosphor sheet comprising: a stimulable phosphor sheet, characterized in that the surface of the protective film is rendered hydrophilic; 2. The stimulable phosphor sheet according to claim 1, wherein the protective film is made hydrophilic by being subjected to a hydrophilic treatment. The stimulable phosphor sheet according to claim 1, characterized in that a hydrophilic layer is provided on the 3° protective film to make it wood-philic. 4. The stimulable phosphor according to any one of claims 1 to 3, wherein the stimulable phosphor is a divalent europium-activated alkaline earth metal fluorohalide phosphor. phosphor sheet.